Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis
Lymphostatin is a virulence factor of enteropathogenic E. coli (EPEC) and non-O157 serogroup enterohaemorrhagic E. coli. Previous studies using whole-cell lysates of EPEC showed that lymphostatin inhibits the mitogen-activated proliferation of bulk human peripheral blood mononuclear cells (PBMCs) an...
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Frontiers Media S.A.
2022-07-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.941939/full |
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| author | Nattaya Ruamsap Nattaya Ruamsap Donporn Riyapa Sujintana Janesomboon Joanne M. Stevens Sathit Pichyangkul Kovit Pattanapanyasat Samandra T. Demons Mark P. Stevens Sunee Korbsrisate |
| author_facet | Nattaya Ruamsap Nattaya Ruamsap Donporn Riyapa Sujintana Janesomboon Joanne M. Stevens Sathit Pichyangkul Kovit Pattanapanyasat Samandra T. Demons Mark P. Stevens Sunee Korbsrisate |
| author_sort | Nattaya Ruamsap |
| collection | DOAJ |
| description | Lymphostatin is a virulence factor of enteropathogenic E. coli (EPEC) and non-O157 serogroup enterohaemorrhagic E. coli. Previous studies using whole-cell lysates of EPEC showed that lymphostatin inhibits the mitogen-activated proliferation of bulk human peripheral blood mononuclear cells (PBMCs) and the production of cytokines IL-2, IL-4, IL-5, and IFN-γ. Here, we used highly purified lymphostatin and PBMC-derived T cells to show that lymphostatin inhibits anti-CD3/anti-CD28-activated proliferation of human CD4+ and CD8+ T cells and blocks the synthesis of IL-2, IL-4, IL-10 and IFN-γ without affecting cell viability and in a manner dependent on an N-terminal DTD glycosyltransferase motif. Such inhibition was not observed with T cells activated by phorbol 12-myristate 13-acetate and ionomycin, implying that lymphostatin targets T cell receptor signaling. Analysis of the expression of CD69 indicated that lymphostatin suppresses T cell activation at an early stage and no impacts on apoptosis or necrosis were observed. Flow cytometric analysis of the DNA content of lymphostatin-treated CD4+ and CD8+ T cells showed a concentration- and DTD-dependent accumulation of the cells in the G0/G1 phase of the cell cycle, and corresponding reduction of the percentage of cells in S phase. Consistent with this, we found a marked reduction in the abundance of cyclins D3, E and A and loss of phosphorylated Rb over time in activated T cells from 8 donors treated with lymphostatin. Moreover, the cyclin-dependent kinase (cdk) inhibitor p27kip1, which inhibits progression of the cell cycle at G1 by acting on cyclin E-cdk2 or cyclin D-cdk4 complexes, was found to be accumulated in lymphostatin-treated T cells. Analysis of the abundance of phosphorylated kinases involved in signal transduction found that 30 of 39 were reduced in abundance following lymphostatin treatment of T cells from 5 donors, albeit not significantly so. Our data provide novel insights into the mode of action of lymphostatin on human T lymphocytes. |
| first_indexed | 2024-12-11T16:26:24Z |
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| language | English |
| last_indexed | 2024-12-11T16:26:24Z |
| publishDate | 2022-07-01 |
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| spelling | doaj.art-2947e196683c47828a49d1783b0341502022-12-22T00:58:43ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-07-011210.3389/fcimb.2022.941939941939Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosisNattaya Ruamsap0Nattaya Ruamsap1Donporn Riyapa2Sujintana Janesomboon3Joanne M. Stevens4Sathit Pichyangkul5Kovit Pattanapanyasat6Samandra T. Demons7Mark P. Stevens8Sunee Korbsrisate9Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandDepartment of Bacterial and Parasitic Diseases, Armed Forces Research Institute of Medical Sciences, Bangkok, ThailandCenter for Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, ThailandDepartment of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Edinburgh, United KingdomDepartment of Bacterial and Parasitic Diseases, Armed Forces Research Institute of Medical Sciences, Bangkok, ThailandDepartment for Research and Development, Siriraj Center of Research Excellence for Microparticle and Exosome in Diseases, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandDepartment of Bacterial and Parasitic Diseases, Armed Forces Research Institute of Medical Sciences, Bangkok, ThailandThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Edinburgh, United KingdomDepartment of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandLymphostatin is a virulence factor of enteropathogenic E. coli (EPEC) and non-O157 serogroup enterohaemorrhagic E. coli. Previous studies using whole-cell lysates of EPEC showed that lymphostatin inhibits the mitogen-activated proliferation of bulk human peripheral blood mononuclear cells (PBMCs) and the production of cytokines IL-2, IL-4, IL-5, and IFN-γ. Here, we used highly purified lymphostatin and PBMC-derived T cells to show that lymphostatin inhibits anti-CD3/anti-CD28-activated proliferation of human CD4+ and CD8+ T cells and blocks the synthesis of IL-2, IL-4, IL-10 and IFN-γ without affecting cell viability and in a manner dependent on an N-terminal DTD glycosyltransferase motif. Such inhibition was not observed with T cells activated by phorbol 12-myristate 13-acetate and ionomycin, implying that lymphostatin targets T cell receptor signaling. Analysis of the expression of CD69 indicated that lymphostatin suppresses T cell activation at an early stage and no impacts on apoptosis or necrosis were observed. Flow cytometric analysis of the DNA content of lymphostatin-treated CD4+ and CD8+ T cells showed a concentration- and DTD-dependent accumulation of the cells in the G0/G1 phase of the cell cycle, and corresponding reduction of the percentage of cells in S phase. Consistent with this, we found a marked reduction in the abundance of cyclins D3, E and A and loss of phosphorylated Rb over time in activated T cells from 8 donors treated with lymphostatin. Moreover, the cyclin-dependent kinase (cdk) inhibitor p27kip1, which inhibits progression of the cell cycle at G1 by acting on cyclin E-cdk2 or cyclin D-cdk4 complexes, was found to be accumulated in lymphostatin-treated T cells. Analysis of the abundance of phosphorylated kinases involved in signal transduction found that 30 of 39 were reduced in abundance following lymphostatin treatment of T cells from 5 donors, albeit not significantly so. Our data provide novel insights into the mode of action of lymphostatin on human T lymphocytes.https://www.frontiersin.org/articles/10.3389/fcimb.2022.941939/fulllymphostatinenteropathogenic Escherichia coliinhibit T cell proliferationcytokine suppressionG0/G1 cell cycle arrestcyclin expression |
| spellingShingle | Nattaya Ruamsap Nattaya Ruamsap Donporn Riyapa Sujintana Janesomboon Joanne M. Stevens Sathit Pichyangkul Kovit Pattanapanyasat Samandra T. Demons Mark P. Stevens Sunee Korbsrisate Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis Frontiers in Cellular and Infection Microbiology lymphostatin enteropathogenic Escherichia coli inhibit T cell proliferation cytokine suppression G0/G1 cell cycle arrest cyclin expression |
| title | Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| title_full | Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| title_fullStr | Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| title_full_unstemmed | Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| title_short | Lymphostatin, a virulence factor of attaching and effacing Escherichia coli, inhibits proliferation and cytokine responses of human T cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| title_sort | lymphostatin a virulence factor of attaching and effacing escherichia coli inhibits proliferation and cytokine responses of human t cells in a manner associated with cell cycle arrest but not apoptosis or necrosis |
| topic | lymphostatin enteropathogenic Escherichia coli inhibit T cell proliferation cytokine suppression G0/G1 cell cycle arrest cyclin expression |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.941939/full |
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